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ysu1310473424.pdf (1.91 MB)
ETD Abstract Container
Abstract Header
Evaluation of the Role of gsp, ygiC, and yjfC Genes in Glutathione Metabolism in Escherichia coli by Gene Disruption
Author Info
Warren, John C., III
Permalink:
http://rave.ohiolink.edu/etdc/view?acc_num=ysu1310473424
Abstract Details
Year and Degree
2011, Master of Science in Chemistry, Youngstown State University, Department of Chemistry.
Abstract
Glutathione metabolism differs amongst humans and bacteria and has the potential to be a good target for antibacterial agents. Within
Escherichia coli
, glutathione can form an amide bond with spermidine to produce the conjugate glutathionylspermidine. This reaction is catalyzed by the enzyme glutathionylspermidine synthetase/amidase. Three genes,
gsp
,
ygiC
and
yjfC
have previously been identified and proposed to be associated with glutathionylspermidine metabolism; however, the specific function of
ygiC
and
yjfC
has yet to be determined. The objective of this research is to eliminate these genes from the
E. coli
genome and evaluate the responsiveness of the knockout strains to various stress conditions. Corresponding genes were replaced by antibiotic resistance genes through homologous recombination. Once the gene disruptions had been achieved, the antibiotic resistance gene was eliminated utilizing a helper plasmid pCP20. The techniques used to create gene disruptions, or knockouts, included polymerase chain reaction and electroporation. Seven strains have been created: single knockout (
gsp
,
ygiC
, or
yjfC
), double knockout (
gsp/ygiC
,
gsp/yjfC
, or
ygiC/yjfC
), and triple knockout (
gsp/ygiC/yjfC
). The sensitivity of the knockout strains were tested by stressors including antibiotics, transition metals, and hydrogen peroxide.
Committee
Nina Stourman, PhD (Advisor)
Jonathan Caguiat, PhD (Committee Member)
Michael Serra, PhD (Committee Member)
Pages
57 p.
Subject Headings
Chemistry
Keywords
Gene disruption
;
Glutathione
;
Spermidine
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Citations
Warren, III, J. C. (2011).
Evaluation of the Role of gsp, ygiC, and yjfC Genes in Glutathione Metabolism in Escherichia coli by Gene Disruption
[Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1310473424
APA Style (7th edition)
Warren, III, John.
Evaluation of the Role of gsp, ygiC, and yjfC Genes in Glutathione Metabolism in Escherichia coli by Gene Disruption.
2011. Youngstown State University, Master's thesis.
OhioLINK Electronic Theses and Dissertations Center
, http://rave.ohiolink.edu/etdc/view?acc_num=ysu1310473424.
MLA Style (8th edition)
Warren, III, John. "Evaluation of the Role of gsp, ygiC, and yjfC Genes in Glutathione Metabolism in Escherichia coli by Gene Disruption." Master's thesis, Youngstown State University, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1310473424
Chicago Manual of Style (17th edition)
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Document number:
ysu1310473424
Download Count:
430
Copyright Info
© 2011, all rights reserved.
This open access ETD is published by Youngstown State University and OhioLINK.